1. Field of the Invention
The present invention relates to the uses of kinsenoside for inhibiting the activation of macrophages, inhibiting the formation of osteoclasts, inhibiting the function of osteoclasts, and/or activating osteoblasts.
2. Descriptions of the Related Art
Bones mainly consist of organic components (e.g. collagenic fibers and mucopolysaccharides), inorganic components (calcium salts and phosphoric salts), water, etc. Bone tissue is in a dynamic equilibrium state. Bone formation and bone resorption, both of which make up the process of bone remodeling, occur in the bone tissue continuously, and they not only can cure slight trauma, but can also enhance the pressure resistance of bones. In addition, the bone formation includes the formation of a new bone matrix and the mineralization of bones.
The bone modeling process relies on the cooperation of two different kinds of cells, osteoblasts and osteoclasts, responsible for the bone formation and the bone resorption, respectively. If any mistake occurs in the coordination between the two cells, it will lead to an imbalance of bone remodeling. For example, if the level of bone resorption is greater than that of bone formation, osteoporosis, commonly seen in clinical medicine (especially postmenopausal women), will take place. On the contrary (i.e. the level of the bone resorption is less than that of the bone formation) is rare, and it may cause an abnormal increase of bone tissue.
Currently, there are about two hundred millions of females with osteoporosis. In 2003, the global market for osteoporosis and hormone supplement therapy for osteoporosis is worth about 8.3 billions USD, and is predicted to reach 17.9 billions USD in 2014. According to mechanisms, pharmaceuticals for osteoporosis can be generally classified into four groups. The first group inhibits the bone resorption, one example of which is diphosphates. The second group stimulates the bone formation, one example of which is parathyroid hormone. The third group inhibits the release of calcium from bones, one example of which is estrogen. The fourth group stimulates the small intestine to absorb calcium, one example of which is Vitamin D. However, diphosphates may bring strong side effects (such as headaches, nausea, vomiting, diarrhea, fever, renal failure, oesophagitis, mandible necrosis, etc). The parathyroid hormone may cause uncomfortableness (such as headaches and nausea). The estrogen has a risk of causing cancer. In addition, the effect of using Vitamin D to enhance the absorption of calcium to improve osteoporosis is quite limited. Therefore, a substance or a pharmaceutical composition that can cure osteoporosis efficiently and with low side effects is still needed.
Macrophages present various differentiated cell forms in different tissues, for instance, Kupffer cells in the liver or microglials in the brain. In the bone tissue, macrophages present a differentiated form as osteoclasts, which maintain a dynamic equilibrium state with osteoblasts.
In the immunological defense mechanism of humans, macrophages in the blood are the first line of defense, and thus, play a very important role in the inflammatory reaction. Specifically, when contacting pathogens, macrophages are activated to trigger the ability for destroying pathogens, and meanwhile secrete cytokines, for example tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), that stimulate the inflammatory reaction. Hence, if the stimulation from pathogens is too strong, or macrophages release too much cytokines within a short period of time, septic shock may arise.
During the inflammatory process, macrophages not only release cytokines, but also present antigens. More specifically, the action of presenting antigens of macrophages may influence the differentiation of T cells, and further influence the generation of B cells and antibodies. B cells and antibodies can also destroy pathogens. Therefore, besides being the first immunological response, macrophages play a role in immunological modulation (see Fuijwara N. Kobayashi K. Macrophage and inflammation. Curr Drug Targets 4, 281-286).
Present studies show that nearly all chronic diseases are relevant to the inflammatory reaction, such as cancers, rheumatic arthritis, etc. In other words, macrophages participate in the morbific mechanisms of these diseases (see Naito M. 2008. Macrophage differentiation and function in health and disease. Pathol Int 58, 143-156).
For rheumatic arthritis, macrophages play an important role in the pathology of this disease. Generally speaking, rheumatic arthritis may activate macrophages to release cytokines, and thus stimulates chronic inflammation. In addition, various kinds of inflammatory cells exist in the joint synovial fluid, and cytokines and growth factors released from the inflammatory cells can make macrophages differentiate into osteoclasts, thus causing damage to joint bones. The main pharmaceutical for rheumatic arthritis currently is steroids. However, steroids have many side effects, such as edema, osteoporosis, etc. Accordingly, a substance or a pharmaceutical composition that can cure rheumatic arthritis efficiently with low side effects is still highly demanded.
Anoectochilus spp. belongs to orchidaceae, and it is believed that Anoectochilus formosanus Hayata has the broad effects of decreasing blood pressure, reducing blood sugar, protecting the liver, anti-inflammation, modulating immune system, and so on. Thus, Anoectochilus formosanus Hayata is also called “the king of drugs” or “the tiger of drugs.” Furthermore, it has been confirmed by documents that the crude extract of Anoectochilus formosanus Hayata has the pharmacological effect of anti-osteoporosis (see Shih C C, Wu Y W, Lin W C. 2001. Ameliorative effects of Anoectochilus formosanus extract on osteopenia in overiectomized rats. J Ethnopharmacol 77, 233-238 and Masuda K, Ikeuchi M, koyama T, Yamaguchi K, Woo J T, Nishimura T, Yazawa K. 2008. Suppressive effects of Anoectochilus formosanus extract on osteoclast formation in vitro and bone resorption in vivo. J Bone Miner Metab 26, 123-129).
Nevertheless, the active component for the anti-inflammation and anti-osteoporosis effects of Anoectochilus formosanus Hayata remains unclear at present, and the optimization of drug efficiency and the pharmacological study are limited accordingly. Moreover, because the traditional suckering proliferation method is quite slow, and the consumption due to collection is not limited, the number of Anoectochilus formosanus Hayata in its natural habitat has decreased dramatically. Although the industry has actively studied cultural methods for Anoectochilus formosanus Hayata (e.g., a vegetative propagation method using the tissue culture of Anoectochilus formosanus Hayata, which can be seen in Shiau Y J, Sagare A P, Chen U C, Yang S R, Tsay H S. 2002. Conservation of Anoectochilus formosanus HAYATA by artificial cross-pollination and in vitro culture of seeds. Bot Bull Acad Sin 43: 123-130, an improved seedling root culture method disclosed in JP 10-056875 A, etc), the number of Anoectochilus formosanus Hayata that can be provided is still restricted by the reproduction/culture speed. Thus, if the active component for anti-inflammation and anti-osteoporosis effects provided by Anoectochilus formosanus Hayata can be obtained, a pharmaceutical composition comprising the active component can be provided and produced in large scale by an artificial synthesis process.
The present invention is the investigation for the above requirements. The inventors of the present invention discovered the main active compound for anti-inflammation and anti-osteoporosis in Anoectochilus formosanus Hayata through related in vivo and in vitro experiments. The active compound has the functions of stimulating the bone formation and inhibiting the bone resorption, and also has the effect of inhibiting the activation of macrophages.